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Beta-glucan-stimulated mammalian myeloid cells, such as macrophages, show an increased responsiveness to secondary stimulation in a nonspecific manner. This phenomenon is known as trained innate immunity and is important to prevent reinfections. Trained innate immunity seems to be an evolutionary conserved phenomenon among plants, invertebrates and mammalian species. Our study aimed to explore the training of primary chicken monocytes. We hypothesized that primary chicken monocytes, similar to their mammalian counterparts, can be trained with β-glucan resulting in increased responses of these cells to a secondary stimulus. Primary blood monocytes of white leghorn chickens were primary stimulated with β-glucan microparticulates (M-βG), lipopolysaccharide (LPS), recombinant chicken interleukin-4 (IL-4) or combinations of these components for 48 h. On day 6, the primary stimulated cells were secondary stimulated with LPS. Nitric oxide (NO) production levels were measured as an indicator of pro-inflammatory activity. In addition, the cells were analyzed by flow cytometry to characterize the population of trained cells and to investigate the expression of surface markers associated with activation. After the secondary LPS stimulation, surface expression of colony stimulating factor 1 receptor (CSF1R) and the activation markers CD40 and major histocompatibility complex class II (MHC-II) was higher on macrophages that were trained with a combination of M-βG and IL-4 compared to unstimulated cells. This increased expression was paralleled by enhanced NO production. In conclusion, this study showed that trained innate immunity can be induced in primary chicken monocytes with β-glucan, which is in line with previous experiments in mammalian species. Innate immune training may have the potential to improve health and vaccination strategies within the poultry sector. [ABSTRACT FROM AUTHOR]

Enhanced dietary Ca intake linearly increases intestinal Ca absorption in pigs, but not in broilers, suggesting potential differences in whole body Ca homeostasis. To determine the role of kidney in Ca homeostasis in these species, we varied in growing pigs in experiment ( Exp ) 1, the dietary Ca content 2.0 vs. 9.6 g/kg and phytase 0 vs. 500 FTU/kg, in broilers, in Exp 2 the dietary Ca/retainable P from 1.3 to 2.8 and phytase 0 vs. 1000 FTU/kg, and in Exp 3 dietary Ca/P from 0.50 to 1.75. Increasing dietary Ca reduced renal mRNA expression of Ca-related transporters (TRPV5, TRPV6, CaBP-D28k, NCX1) and tight junctions (CLDN-12 and -16) in pigs, indicating Ca reabsorption was reduced to maintain Ca homeostasis. In broilers (Exp 2), high dietary Ca increased renal TRPV6, CaBP-D28k and CLDN-2 mRNA, indicating an increased capacity for Ca reabsorption. Moreover, the effect of dietary Ca was enhanced by inclusion of dietary phytase in pigs but reduced in broilers. Furthermore, increasing dietary Ca upregulated PiT-1 while phytase downregulated XPR1 mRNA expression in pigs; in broilers dietary Ca downregulated renal mRNA expression of NaPi-IIa, PiT-1, PiT-2 and XPR1, while phytase downregulated NaPi-IIa but upregulated PiT-2 and XPR1 mRNA expression. In Exp 3, Ca/P effect on transporter mRNA expression was largely consistent with Exp 2. In conclusion of this study, together with previously measured data about Ca and P homeostasis, in pigs the kidneys play a more regulatory role in Ca homeostasis than in broilers where the intestine is more important for regulation.

The present study aimed to confirm the previously reported 'recovery' effect to a challenging diet (CD) of a Lactobacilli-based probiotic (Pro) and its derived postbiotic (Post) in broilers. Identical diet compositions were used, and observations were extended to a second CD diet. A completely randomised block design of 2 × 3 treatment groups with two CDs and three additive conditions (Control, Pro, Post) was used. One additional group received a standard diet (SD). The study involved 1600 one-day-old Ross 308 male broilers. All diets, fed from d1 to 35, were formulated to contain identical nutrients levels, with CDs formulated to be greater than SD in nonstarch polysaccharides using rye and barley (Rye CD) or dry distiller grains with soluble, sunflower and rapeseed meal (DDGS CD). Growth performance parameters, footpad lesions (FPL) score and plasma Ca, P and uric acid concentrations were measured. Compared to SD, birds fed Rye CD and DDGS CD had a higher 1-35 days feed conversion ratio (+3.4 and +4.1%, respectively), due to a higher feed intake for Rye CD (+2.9%) and a lower body weight for DDGS CD (-4.1%). An effect of additive was restricted to Rye CD where Post depressed BW at d28 and d35 (-3.7 and -2.4%, respectively). Compared to Rye CD, DDGS CD lowered plasma Ca/P at d21 (-9.0%) and d35 (-8.1%) and uric acid at d21 (-26%). Pro increased plasma Ca in Rye CD at d21 (+12%) and Post decreased plasma uric acid in DDGS CD at d35 (-25%). All other plasma parameters were not affected. The previously observed recovery effect of a commercial probiotic and postbiotic were not reproducible under highly similar growth conditions, which suggests that both may have specific physiological effects which are only expressed under specific circumstances., (© 2024 The Author(s). Journal of Animal Physiology and Animal Nutrition published by Wiley‐VCH GmbH.)

Diacylglycerol kinase (DGK) phosphorylates the second messenger diacylglycerol (DAG) to phosphatidic acid. We previously identified DGK as one of nine mammalian DGK isoforms and reported on its regulation by interaction with RhoA and by translocation to the plasma membrane in response to noradrenaline. Here, we have investigated how the localization of DGK, fused to green fluorescent protein, is controlled upon activation of G protein-coupled receptors in A431 cells. Extracellular ATP, bradykinin, or thrombin induced DGK translocation from the cytoplasm to the plasma membrane within 2-6 min. This translocation, independent of DGK activity, was preceded by protein kinase C (PKC) translocation and was blocked by PKC inhibitors. Conversely, activation of PKC by 12-O-tetradecanoylphorbol-13-acetate induced DGK translocation. Membrane-permeable DAG (dioctanoylglycerol) also induced DGK translocation but in a PKC (staurosporin)-independent fashion. Mutations in the cysteine-rich domains of DGK abrogated its hormone- and DAG-induced translocation, suggesting that these domains are essential for DAG binding and DGK recruitment to the membrane. We show that DGK interacts selectively with and is phosphorylated by PKCepsilon and -eta and that peptide agonist-induced selective activation of PKCepsilon directly leads to DGK translocation. Our data are consistent with the concept that hormone-induced PKC activation regulates the intracellular localization of DGK, which may be important in the negative regulation of PKCepsilon and/or PKCeta activity.

Diacylglycerol kinase (DGK) phosphorylates the second messenger diacylglycerol (DAG) to phosphatidic acid (PA). Among the nine mammalian isotypes identified, DGKtheta is the only one with three cysteine-rich domains (CRDs) (instead of two) in its N-terminal regulatory region. We previously reported that DGKtheta binds to and is negatively regulated by active RhoA. We now report that RhoA strongly binds to the C-terminal catalytic domain, which would explain its inhibition of DGK activity. To help finding a physiological function of DGKtheta, we further determined its activity in vitro as a function of 15 different truncations and point mutations in the primary structure. Most of these alterations, located throughout the protein, inactivated the enzyme, suggesting that catalytic activity depends on all of its conserved domains. The most C-terminal CRD is elongated with a stretch of 15 amino acids that is highly conserved among DGK isotypes. Mutation analysis revealed a number of residues in this region that were essential for enzyme activity. We suggest that this CRD extension plays an essential role in the correct folding of the protein and/or in substrate presentation to the catalytic region of the protein.